TW202216182A - Use of phellinus linteus for preparing composition of improving sarcopenia - Google Patents

Abstract

The invention provides a use of Phellinus linteus for preparing composition of improving sarcopenia, in which the Phellinus linteus can be the strain with a deposit number of BCRC 930210. The aforementioned composition includes an effective substance, and the aforementioned effective substance is oriented from a fermented extract of Phellinus linteus and/or derivative of the same. By using the aforementioned composition including the fermented extract and/or its derivative, diameters of myotubes, amounts of muscles and muscle endurance can be maintained, thereby improving sarcopenia.

Description

Use of Phellinus linteus for preparing the composition for improving sarcopenia

The present invention relates to the use of a Phellinus linteus, in particular to the use of a Phellinus linteus for preparing a composition for improving sarcopenia.

Sarcopenia is one of the common diseases in old age. The characteristics of sarcopenia include the continuous reduction of systemic skeletal muscle weight and function. If sarcopenia continues to deteriorate, it will not only cause the patient's activity and quality of life to decline, but may also increase the incidence of other diseases, disability, and falls. even lead to death.

Causes of sarcopenia include motor nerve degeneration, nutrient imbalances, decreased protein synthesis, chronic diseases and/or inflammatory reactions. At present, sarcopenia cannot be controlled by drugs, so exercise, control of chronic diseases, inflammatory symptoms and supplementation of specific nutrients are needed to delay or improve sarcopenia.

Phellinus linteus, also known as mulberry and mulberry, is a medicinal fungus belonging to the genus Phellinus in the family Embroideryaceae. Phellinus grows on tree trunks, especially those of mulberry plants. Phellinus linteus is low in toxicity but has many effects, for example, antioxidant, anti-inflammatory, immune-boosting, anti-cancer, liver-protecting, anti-dementia, cardiovascular disease prevention, anti-allergic (allergic rhinitis, eczema, Rheumatoid arthritis), sleep relief, analgesia (such as menstrual pain), inhibition of uric acid and skin care, etc. However, there are few studies on the efficacy of Phellinus linteus in improving sarcopenia.

Therefore, one aspect of the present invention is to provide the use of Phellinus linteus for preparing a composition for improving sarcopenia, so as to maintain myotube diameter, muscle mass and muscular endurance.

According to the above aspect of the present invention, there is provided a use of Phellinus linteus for preparing a composition for improving sarcopenia, wherein the registration number of Phellinus linteus is BCRC 930210. The above-mentioned composition contains an effective substance, and the effective substance can be derived from, for example, the fermented extract of Phellinus linteus and/or its derivatives.

According to the above-mentioned embodiment of the present invention, the above-mentioned fermented extract can be obtained, for example, by performing a multi-stage culturing step and an extraction step on the first mycelium of Phellinus linteus. First, the first mycelium is subjected to a solid-state culture step using a solid medium at 15°C to 30°C for 1 to 2 weeks to obtain a second mycelium. Then, at 15 DEG C to 30 DEG C, utilize the first culture solution to carry out liquid culture step to the second mycelium for 3 to 14 days to obtain the third mycelium, wherein the pH value of the first culture solution is pH 2 to pH 6. Next, at 15 DEG C to 30 DEG C, utilize the second nutrient solution to carry out fermentation culture step to the third mycelium for 3 to 21 days, to obtain fermented product, wherein the pH value of the second nutrient solution is pH 2 to pH 6.

According to the above-mentioned embodiment of the present invention, the fermented extract comprises fermented water extract and/or fermented wine extract, and the derivative can be selected from, for example, a dried fermented water extract, a fermented water extract concentrate, A group consisting of the dried fermented wine extract, the concentrated fermented wine extract and any combination of the above.

According to the above-mentioned embodiment of the present invention, the fermented water extract is obtained by water extraction, and the water extraction comprises performing hot water extraction of the fermented product with water at 100°C.

According to the above-mentioned embodiment of the present invention, the fermented wine extract is obtained by wine extraction, and the wine extraction includes ultrasonically vibrating the fermented product with ethanol.

According to the above-mentioned embodiment of the present invention, the effective dose of the fermented water extract on animal muscle cells can be, for example, 5 μg/mL to 15 μg/mL.

According to the above-mentioned embodiment of the present invention, the effective dose of the fermented wine extract on animal muscle cells can be, for example, 0.5 μg/mL to 1.5 μg/mL.

According to the above-mentioned embodiment of the present invention, when the composition is administered to mice, the effective dose of the effective substance can be, for example, 400 mg/kg.body weight (bw)/day to 600 mg/kg.bw/day.

According to the above-mentioned embodiment of the present invention, when the composition is administered to the human body, the effective dose of the active substance can be, for example, 2300 mg/60 kg.bw/day to 2500 mg/60 kg.bw/day.

According to the above-mentioned embodiments of the present invention, the composition can be, for example, a pharmaceutical composition or a food composition, and the composition can include, but is not limited to, food or medically acceptable carriers, excipients, diluents, adjuvants, preservatives, fillers and/or additives.

Apply the Phellinus linteus of the present invention to prepare a composition for improving sarcopenia, the composition contains the fermented extract of Phellinus linteus and/or its derivatives as effective substances, and is administered to the subject to maintain muscle Tube diameter, muscle mass and muscular endurance.

Based on the above, the present invention provides a use of Phellinus linteus for preparing a composition for improving sarcopenia, wherein the composition comprises an effective substance, and the effective substance can be derived from, for example, a fermented extract of Phellinus linteus and/or derivatives thereof.

The above-mentioned Phellinus linteus can be deposited in the Biological Resource Center (BCRC) of the Food Industry Development Research Institute, No. 331, Food Industry Road, Hsinchu, Taiwan, for example, on July 18, 2019, and the deposit number is the bacterial strain of BCRC 930210, wherein Phellinus linteus is mycelium. form of storage.

After the above-mentioned Phellinus Phellinus mycelium is subjected to a multi-stage culture step, the fermentation product of Phellinus Phellinus can be obtained. Since Phellinus linteus requires different culture conditions (including: nutrients and environmental factors) in different growth or differentiation stages, it is necessary to adjust the culture conditions of Phellinus Phellinus in each growth stage through multi-stage culture steps to obtain more biological amount and/or specific active ingredients.

In one embodiment, the multi-stage culturing step may include a solid-state culturing step, a liquid culturing step, and a fermenting culturing step. Specifically, in the solid-state culture step, for example, the above-mentioned mycelium (or the first mycelium) of Phellinus linteus can be cultured in a solid medium to obtain the second mycelium. The above-mentioned solid medium can contain carbon source, nitrogen source and other nutrients required for the growth of Phellinus linteus. In one embodiment, the solid medium may be, for example, potato dextrose agar (PDA). In one embodiment, the solid state culturing step may be, for example, culturing at 15°C to 30°C for 1 week to 2 weeks.

The above-mentioned liquid culturing step can, for example, use the first nutrient solution to perform a liquid culturing step on the second mycelium to obtain the third mycelium, wherein the pH value of the first nutrient solution can be, for example, pH 2 to pH 6, and the first The culture medium can comprise 1% to 3% by weight of comprehensive carbon and nitrogen sources (such as cereals and/or beans), 1% to 4% by weight of carbohydrates (such as monosaccharides and/or disaccharides), 0.1% by weight to 1 wt% yeast extract, 0.1 wt% to 1 wt% protein gluten and 0.01 wt% to 0.05 wt% inorganic salts (eg phosphate and/or sulfate). It should be understood that, the composition of the aforementioned first culture solution can be appropriately adjusted according to the needs of use. In one embodiment, the first culture solution can be cultured at 15°C to 30°C with a rotation speed of 110 rpm to 130 rpm for 3 days to 14 days.

The above-mentioned fermentation culture step can for example utilize the second culture solution to carry out the fermentation culture step to the third mycelium for 3 days to 21 days at 15 ° C to 30 ° C to obtain the fermentation product, wherein the second culture solution is The composition of the first medium can be, for example, the same as that of the first medium, or its composition can be appropriately adjusted according to the needs of use, and the pH value of the second medium can be, for example, 2 to 6.

The above-mentioned fermentation step is carried out in a fermentation tank. In one embodiment, during the fermentation culturing step, gas is introduced into the fermentation tank, wherein the gas can be selected from a group consisting of air, oxygen, carbon dioxide, helium and any combination thereof. In one embodiment, the cell pressure may be, for example, 0.5 kg/cm ² to 1.0 kg/cm ² . In one embodiment, the aeration rate may be, for example, 0.01 (volume of gas vented/volume of broth per minute, VVM) to 1.5 VVM. In other embodiments, the rotational speed of the fermentation step is 50 rpm to 150 rpm.

Next, the above-mentioned fermented substance is subjected to an extraction step to obtain a fermented fermented substance. The extraction step can be performed using conventional extraction methods. In one embodiment, the fermented product is subjected to solvent extraction with a polar solvent, wherein the polar solvent includes water and/or lower alcohols (eg, methanol, ethanol, propanol, isopropanol, etc.). It is worth noting that, considering the needs of subsequent applications, it is better to use water or ethanol for water extraction or wine extraction, respectively, to obtain fermented water extract or fermented wine extract.

In one embodiment, the above-mentioned water extraction can be selectively performed with boiling water (eg, at 90°C to 100°C) for hot water extraction. In another embodiment, the above-mentioned wine extraction can be selectively combined with ultrasonic vibration treatment, so as to use ultrasonic waves of, for example, 500 W to 700 W, 30 kHz to 50 kHz at room temperature (for example, 10° C. to 40° C.). . The time of the above-mentioned extraction step is not limited. In one embodiment, the hot water extraction is performed for 20 to 40 minutes. In another embodiment, the above-mentioned wine extraction combined with ultrasonic vibration treatment is carried out for 40 minutes to 80 minutes to obtain effective substances with a higher content.

In the above embodiment, the ratio of the polar solvent to the weight of the leaven is not particularly limited, and can be, for example, 10 times to 30 times, but not limited to the ones listed here.

In one embodiment, between the multi-stage culturing step and the extraction step, reprocessing may be included but not limited to, wherein the reprocessing may include a drying step and/or a concentration step to facilitate the subsequent extraction step.

The above drying step can be carried out by conventional drying methods, such as freeze drying, vacuum drying or spray drying. In one embodiment, a drying step is performed on the fermented product to obtain a dried fermented product.

The above-mentioned concentration step can be carried out by conventional concentration methods, such as concentration under reduced pressure, evaporation concentration method or membrane concentration method.

In one embodiment, after the extraction step, the above-mentioned reprocessing may also be included, but not limited to, to obtain a derivative of the fermented extract. The derivative can be selected from the group consisting of dried fermented water extract, fermented water extract concentrate, dried fermented wine extract, fermented wine extract concentrate and any combination thereof.

The above-mentioned fermented extract and/or its derivatives have been confirmed to be useful for preparing a composition for improving sarcopenia. The "effect of improving sarcopenia" means that after the above-mentioned composition is administered to the subject, it can delay the loss of muscle mass and muscle endurance, and the specific evaluation indicators include the ability to effectively maintain the diameter of myotubes, muscle mass and muscle endurance. endurance.

It has been confirmed by animal cell experiments that fermented water extract and/or fermented wine extract can maintain the effect of myotube diameter. In this embodiment, the effective dose of fermented water extract to animal muscle cells can be, for example, 5 μg/mL to 15 μg/mL, and the effective dose of fermented wine extract to animal muscle cells can be, for example, 0.5 μg/mL mL to 1.5 μg/mL. The above dosage range is sufficient for the fermented water extract and/or the fermented wine extract to have sarcopenia delaying effect without toxicity to muscle cells.

In addition, the fermented water extract and/or the fermented wine extract may have the effect of maintaining muscle mass and muscular endurance after evaluation in animal experiments.

In application, the aforementioned fermented extract and/or its derivatives can be added to the composition, and the effective dose thereof depends on the type of the fermented extract and/or its derivatives and/or the object of administration. In one embodiment, when the fermented water extract and/or the fermented wine extract is administered to mice, the effective dose can be, for example, 400 mg/kg. body weight (body weight, bw)/day to 600 mg/kg. bw/day. In one embodiment, when the fermented water extract and/or the fermented wine extract is administered to the human body, the effective dose may be, for example, 2300 mg/60 kg.bw/day to 2500 mg/60 kg.bw/day.

In application, the aforementioned composition may be, for example, a food composition or a pharmaceutical composition. In one embodiment, the composition may optionally include food or medically acceptable carriers, excipients, diluents, adjuvants, preservatives, fillers and/or additives.

Several embodiments are used below to illustrate the application of the present invention, but they are not intended to limit the present invention. Those with ordinary knowledge in the technical field of the present invention can make various changes and modifications without departing from the spirit and scope of the present invention. retouch.

Embodiment 1. Preparation of fermented extract of Phellinus linteus and derivatives thereof

The first mycelium of Phellinus linteus in this example was deposited in the Biological Resource Research Center (BCRC) of the Food Industry Development Research Institute of a consortium, and the deposit number is the strain 930210. This strain is mycelium isolated from wild Phellinus linteus fruiting bodies in China.

For the mycological characteristics and cultivation methods of the above-mentioned Phellinus linteus (BCRC 930210), please refer to the case of Taiwan Application No. TW109129939, which is incorporated herein by reference.

The mycelium (or the first mycelium) of Phellinus linteus deposited in BCRC was inoculated on potato dextrin medium (potato dextrose agar, PDA), and cultivated at 25°C for 7 days to obtain the second bacteria. silk. Then, scrape part of the second mycelium of Phellinus linteus and inoculate it in the first nutrient solution, to carry out a culturing step for 7 days at 25° C., pH 5, and rotating speed 120 rpm, to obtain the third bacteria Filament, wherein the above-mentioned first culture solution contains 1% by weight of comprehensive carbon and nitrogen sources, 1.5% by weight of carbohydrates, 0.3% by weight of yeast extract, 0.3% by weight of protein hydration and 0.05% by weight of inorganic salts. The above-mentioned comprehensive carbon and nitrogen sources are cereals (wheat flour and/or bran flour) and/or beans (soybean flour, mung bean flour, soybean flour and/or cinnamon flour). The above sugars are monosaccharides (glucose and/or fructose) and/or disaccharides (maltose and/or sucrose). The above-mentioned inorganic salts are phosphates (dipotassium hydrogen phosphate, potassium dihydrogen phosphate) and/or sulfates (magnesium sulfate and/or iron sulfate). The specific formulations of the comprehensive carbon and nitrogen sources, sugars and inorganic salts are well known to those with ordinary knowledge in the technical field to which the present invention pertains, and can be arbitrarily adjusted according to actual needs without affecting the fermentation step, and will not be repeated here.

Then, the third body mycelium of the part of Phellinus linteus in the first nutrient solution is inoculated in the fermenting tank containing the second nutrient solution (the composition is identical to the first nutrient solution), with 25° C., pH 5, 0.5 The fermentation was carried out at an air pressure of kg/cm ² , an air ventilation rate of 1.0 VVM, and a stirring speed of 80 rpm for 14 days to obtain a starter.

Take the leaven and freeze-dry to obtain the freeze-dried powder of the leaven. In this example, 100 L of fermented yeast can obtain 3 kg of fermented lyophilized powder.

Next, an extraction step is performed on the fermented lyophilized powder to obtain a fermented extract, wherein the fermented extract comprises a fermented water extract and a fermented wine extract. Above-mentioned fermented water extract is to utilize distilled water to carry out hot water extraction at 100 DEG C and obtain after 30 minutes, and wherein the weight ratio of water to fermented lyophilized powder is 20. After the fermented water extract was cooled to room temperature, freeze-drying was performed to obtain sample 1.

Above-mentioned fermented wine extract is to utilize ethanol at 25 DEG C to carry out ultrasonic shock treatment and obtain after reaching 1 hour, and wherein ultrasonic shock treatment is to utilize ultrasonic cleaner (manufacturer: Delta Ultrasonic Co., Ltd., Taiwan; Model: DC600H) is carried out with the ultrasonic wave of 600 W, 40 kHz, and the weight ratio of ethanol to fermented lyophilized powder is 20. Next, centrifugation is performed to separate layers, wherein the supernatant is the fermented wine extract. Then, the fermented wine extract was concentrated under reduced pressure to obtain sample 2.

Samples 1 and 2 were dissolved with Dimethyl sulfoxide (DMSO), respectively, to facilitate the configuration described later.

Example 2. Evaluate the effect of fermented ferment extract to maintain myotube diameter, muscle mass and muscle endurance

A sarcopenia model was established using mouse skeletal muscle cells (C2C12 cell line). The sarcopenia model can be established using the synthetic corticosteroid dexamethasone, which suppresses the immune system and thus acts as an anti-inflammatory or anti-allergic drug. However, dexamethasone also has the side effect of causing muscle atrophy (including decreased muscle mass and/or decreased muscular endurance), so this example uses dexamethasone to induce muscle cell atrophy to simulate sarcopenia.

First, use growth medium [Dulbecco's modified minimal essential medium (DMEM) with 10% fetal bovine serum] to culture C2C12 cell line at 37°C, 5% CO ₂ , wherein the starting point of C2C12 cell line is ^The starting cell density was 1x105 to ^2x105 cells/mL. When the C2C12 cell line was 70% full, it was cultured in differentiation medium (DMEM containing 2% horse serum) for 7 days, during which the differentiation medium was changed every 2 days to obtain differentiated cells, of which 90% were differentiated myotubes.

Next, the above-mentioned differentiated cells are divided into blank control group, negative control group, experimental group 1, experimental group 2, experimental group 3 and experimental group 4, wherein the medium of the blank control group is DMEM containing 0.1% DMSO, and the negative control The medium of group 1 was DMEM containing 0.1% DMSO and 10 μM dexamethasone, the medium of experimental group 1 was DMEM containing 10 μg/mL of sample 1, and the medium of experimental group 2 was DMEM containing 10 μM dexamethasone and 10 μg/mL of dexamethasone. mL of DMEM of sample 1, medium of experimental group 3 was DMEM containing 1 μg/mL of sample 2, and medium of experimental group 4 was DMEM of sample 2 containing 10 μM dexamethasone and 1 μg/mL. It is worth noting that because sample 1 and sample 2 were first dissolved in DMSO and then used to prepare the medium, the medium of experimental group 1 to experimental group 4 contained DMSO, but the concentration was less than or equal to 0.1% to avoid causing damage to cells. toxicity.

After culturing the above-mentioned differentiated cell lines in the above-mentioned medium for 24 hours at 37°C and 5% _CO , respectively, the cells were stained with hematoxylin and eosin (H&E), and then the myotubes in the differentiated cell lines were observed with a light microscope. The size of the diameter is shown in Figure 1A to Figure 1F.

FIGS. 1A to 1F are histological staining diagrams of culturing mouse skeletal muscle cells in a medium with or without fermented fermented extract and/or dexamethasone according to an embodiment of the present invention, wherein FIG. 1A , FIG. 1B , FIG. 1C , FIG. 1D , FIG. 1E and FIG. 1F correspond to the blank control group, the negative control group, the experimental group 1, the experimental group 2, the experimental group 3 and the experimental group 4, respectively. Compared with Fig. 1A (blank control group), the diameter of myotubes in Fig. 1B (negative control group) is smaller, but compared with Fig. 1B (negative control group), Fig. 1C to Fig. 1F (experimental group 1 to experimental group 4) ) myotubes are larger in diameter.

The diameter of myotubes was measured by a commercially available software (Image-Pro Plus software), and the statistical results of the diameters of myotubes are shown in Table 1, wherein the statistical method in Table 1 was analyzed by paired sample t-test (paired sample t-test) The percentages of each item, and "#" and "*" represent statistically significant differences with the blank control group and the negative control group (p<0.05, n=60).

Table 1

It can be seen from Table 1 that compared with the blank control group, the diameter of the myotubes in the negative control group decreased significantly, indicating that dexamethasone can indeed cause muscle cell atrophy. However, compared to the negative control group, the experimental group 2 and the experimental group 4 were significantly higher, indicating that the fermented water extract and/or the fermented wine extract could maintain the length of the myotube diameter induced by dexamethasone, which means The fermented water extract and/or the fermented wine extract can delay and/or prevent the loss of muscle mass. In addition, there was no significant difference between the experimental group 1 and experimental group 3 and the blank control group, indicating that the fermented water extract or the fermented wine extract did not affect the size of the diameter of the myotubes, in other words, the fermented water extract or the fermented wine extract The extract was not cytotoxic to myotubes.

Embodiment 3. Evaluate the effect of fermented ferment extract to maintain muscle mass and muscle endurance

The above-mentioned sample 1 and sample 2 were mixed in equal weight to obtain sample 3.

In this example, C57BL/6J mice were used as model organisms. The mice were divided into blank group, control group and experimental group, and were fed with the corresponding reagent once a day, wherein the reagent of the experimental group was sample 3, and the reagent of the blank group and the control group was water. Specifically, sample 3 was dissolved in an appropriate amount of water to control the feeding amount at 500 mg/kg.bw/time, and the volume of water in the blank group and control group was equal to the total volume of sample 3 mixed with water.

The hindlimbs of the mice in the experimental group and the control group were treated with plaster cast for 7 days to induce atrophy of the hindlimbs of the mice. Next, the cast was removed and the mice were allowed to move freely in the rat cage for 7 days. Then, a muscle endurance experiment was performed, and the mice were sacrificed to measure the hindlimb skeletal muscles of the mice. Tube feeding was continued during the above cast and free movement period (14 days in total).

The above-mentioned muscular endurance experiment is to place the mice on an inclined treadmill, wherein the conveyor belt of the treadmill is transported down at a speed of 18 m/min to 20 m/min, and the bottom of the treadmill is provided with a fright grid ( shock grids). If the mouse is not moving, the conveyor belt transports the mouse to the bottom of the treadmill, giving the mouse an electric shock to the tail. In general, mice run up to avoid shocks to their tails. However, if the muscular endurance was insufficient, the mice could not overcome the speed of the conveyor belt and were transported to the bottom of the treadmill, where they received electric shocks. In the same time, if the mice received more electric shocks, the muscle endurance of the mice was worse. Using commercially available software [GraphPad Prism (version 8.0)] to analyze the number of electric shocks received by mice with one-way ANOVA and Dunnett's test post-hoc, the results are shown in Table 2. Among them, "#" and "*" indicate statistically significant differences with blank group and control group (p<0.05, n=6).

Table 2

As shown in Table 2, compared with the blank group, the number of electric shocks received by the mice in the control group was significantly increased, indicating that the plaster immobilization method can indeed cause the skeletal muscle atrophy of the mice and the loss of muscle endurance. However, compared to the control group, the number of electric shocks received by the mice in the experimental group decreased significantly, indicating that administration of a mixture of fermented water extract and fermented wine extract can effectively maintain muscle mass and muscular endurance, thereby delaying and/or Avoid loss of muscular endurance in mice.

Next, the mice were sacrificed and the weight of their hindlimb gastrocnemius muscle was measured. The results were analyzed by one-way ANOVA and Dunnett's test post-hoc analysis using commercially available software [GraphPad Prism (version 8.0)] to count differences between groups. The results are shown in Table 3, in which the relative weight is the weight of the gastrocnemius muscle divided by the weight of the mouse to exclude the factors of individual differences in body size, and the figures "#" and "*" indicate the difference between the blank group and the control group, respectively. Statistically significant difference (p<0.05, n=6).

table 3

As shown in Table 3, compared with the blank group, the relative weight of the gastrocnemius muscle in the control group was significantly smaller, indicating that the plaster immobilization treatment can cause a decrease in muscle mass. However, the relative weight of the gastrocnemius muscle in the experimental group was significantly larger compared to the control group, indicating that administration of a mixture of fermented water extract and fermented wine extract can effectively maintain muscle mass, thereby delaying and/or avoiding muscle mass gain. churn.

Embodiment 4. Estimating the effective dose of human body

In this example, according to the preliminary estimation method of the experiment announced by the US Food and Drug Administration in 2005, the above-mentioned effective dose in mice is used to estimate the effective dose in humans. This method divides the effective dose per kilogram body weight of mice by the conversion factor 12.3, which is the effective dose per kilogram body weight of human body. According to the estimated daily dose of 500 mg/kg.bw administered to mice in this example, the effective dose for humans is 2400 mg/60 kg.bw/day (calculated based on the body weight of an adult being 60 kg.bw).

From the above, it can be seen that the fermented water extract and the fermented wine extract of the present invention can effectively maintain the diameter of myotubes, muscle mass and muscle endurance, and thus can be used to improve sarcopenia.

It should be added that although the present invention takes a specific process, a specific analysis method and/or a specific instrument as an example to illustrate the use of the Phellinus linteus of the present invention for preparing a composition for improving sarcopenia, the present invention belongs to the technical field. Anyone with ordinary knowledge will know that the present invention is not limited to this, without departing from the spirit and scope of the present invention, the use of Phellinus linteus of the present invention for preparing the composition for improving sarcopenia can also use other processes, other analytical methods or other instruments.

Although the present invention has been disclosed above with several embodiments, it is not intended to limit the present invention. Anyone with ordinary knowledge in the technical field to which the present invention belongs, without departing from the spirit and scope of the present invention, can make various Therefore, the scope of protection of the present invention should be determined by the scope of the appended patent application.

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In order to make the above and other objects, features, advantages and embodiments of the present invention more clearly understood, the detailed description of the accompanying drawings is as follows: [FIG. 1A] to [FIG. 1F] show micrographs of tissue staining of mouse skeletal muscle cells cultured in a medium with or without fermented extract and dexamethasone according to one embodiment of the present invention.

The mycelium and/or its derivatives of Phellinus linteus were deposited at the Biological Resource Center (BCRC), Food Industry Development Institute, No. 331, Food Road, Hsinchu, Taiwan on July 18, 2019, and the deposit number is BCRC 930210 strain.

Claims (10)

A kind of Phellinus linteus is used to prepare the purposes of the composition of improving sarcopenia, wherein the deposit number of this Phellinus linteus is BCRC 930210, and this composition comprises an effective substance, and this effective substance is derived from this Phellinus linteus A fermented extract and/or its derivatives. The use of Phellinus linteus as claimed in claim 1 for preparing a composition for improving sarcopenia, wherein the fermented extract is obtained by subjecting one of the first mycelia of Phellinus linteus to a multi-stage culture step and a The extraction step is obtained, wherein the multi-stage cultivation step comprises: At 15 DEG C to 30 DEG C, utilize a solid medium to carry out a solid state culture step to this first mycelium for 1 week to 2 weeks, to obtain a second mycelium; At 15 ℃ to 30 ℃, utilize a first nutrient solution to carry out a liquid culturing step to this second mycelium for 3 to 14 days to obtain a third mycelium, wherein one of the first nutrient solutions pH values ranging from pH 2 to pH 6; and At 15 ℃ to 30 ℃, utilize a second nutrient solution to carry out a fermentation culture step to the third mycelium for 3 days to 21 days, to obtain this fermented product, wherein an acid-base of this second nutrient solution Values are pH 2 to pH 6. The use of Phellinus linteus as claimed in claim 2 for preparing a composition for improving sarcopenia, wherein the fermented extract comprises a fermented water extract and/or a fermented wine extract, and the derivative is It is selected from the group consisting of one fermented water extract dried product, one fermented water extract concentrate, one fermented wine extract dried product, one fermented wine extract concentrate and any combination thereof. The use of Phellinus linteus as claimed in claim 3 for preparing a composition for improving sarcopenia, wherein the fermented water extract is obtained by a water extraction, and the water extraction comprises water to a water at 100° C. The leaven is subjected to a hot water extraction. The use of Phellinus linteus as claimed in claim 3 for preparing a composition for improving sarcopenia, wherein the fermented wine extract is obtained by a wine extraction, and the wine extraction comprises performing a fermentation on a fermented product with ethanol Ultrasonic vibration treatment. The use of Phellinus linteus as claimed in claim 3 for preparing a composition for improving sarcopenia, wherein an effective dose of the fermented water extract to an animal muscle cell is 5 μg/mL to 15 μg/mL. The use of Phellinus linteus according to claim 3 for preparing a composition for improving sarcopenia, wherein an effective dose of the fermented wine extract to an animal muscle cell is 0.5 μg/mL to 1.5 μg/mL. The use of Phellinus linteus as claimed in claim 1 for preparing a composition for improving sarcopenia, wherein when the composition is administered to a mouse, an effective dose of the effective substance is 400 mg/kg. body weight (body wight, bw)/day to 600 mg/kg.bw/day. The use of Phellinus linteus according to claim 1 for preparing a composition for improving sarcopenia, wherein when the composition is administered to a human body, an effective dose of the effective substance is 2300 mg/60 kg.bw/day to 2500 mg/60 kg.bw/day. Use of Phellinus linteus as claimed in claim 1 for preparing a composition for improving sarcopenia, wherein the composition is a pharmaceutical composition or a food composition, and the composition further comprises a food or a medically acceptable A carrier, an excipient, a diluent, an adjuvant, a preservative, a filler and/or an additive are accepted.

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